Bis quaternary oximes



United States Patent 1 Claim. (Cl. 167-65) (Granted under Title 35, US.Code (1952.), sec. 266) This invention described herein may bemanufactured and used by or for the Government of the United States ofAmerica for governmental purposes without the payment to us of anyroyalty thereon.

This is a division of application Serial No. 809,578, filed April 28,1959, now Patent No. 3,077,476, granted March 12, 1963.

This invention is directed to certain diquaternary pyridinium halideoximes which are useful as chemotherapeutic and prophylactic agents formammals poisoned by anticholinesterases, particularly the nerve gasknown as GB or sarin, i.e., isopropyl methylphosphonofluoridate.

The invention relates to 1,1'-polymethylene bis(4- formylpyridinium)halide dioximes wherein the polymethylene group contains from 2 to 6carbon atoms; These compounds have the structural formula The inventionalso relates to 1,1'-(2-'butenylene) bis (4-formylpyridinium) halidedioximes of the formula 0 H=N O H GH=N O E (III) The invention furtherrelates to 1,1'-polymethylene bis (3-formylpyridinium) halide dioximesof the formula wherein R is a polymethylene group containing from 2 to 6carbon atoms.

Another class of compounds included are the 1,1-(pphenylenedimethylene)bis (3-formy1pridinium) halide Patented June 11, 1963 dioximes of theformula i CH==NOH (IVb) This invention further relates to unsymmetricalbisquaternary 4-formylpyridinium halide monoximes of the formula 0 Thisinvention also relates to the compounds This invention also relates tothe compounds CH=NOH (VIII) While all these compounds are useful invarying degree for the purposes set out above, the difierent groupsexhibit quite striking differences in effectiveness. In all the aboveformulas X is chloride, bromide or iodide, which appear to beequivalents as to physiological action, except for their effect onsolubility.

Wilson et -al., in Patent No. 2,816,113, granted December 10, 1957,disclose a group of compounds which are effective as antidotes fortreatment of mammals poisoned by compounds of high anticholinesteraseactivity, such as the nerve gases diisopropylphosphorofluoridate (DFP),isopropyl methylphosphonofluoridate (GB), and O-ethyl, N,N dimethylphosphoroamidocyanidate (GA), as well as other related organicphosphorus compounds, including many insecticides. The compound of theWilson et al. group which has received by far the most attention is2-formyl-l-methyl pyridinium iodide oxime, commonly known as Z-pyridinealdoXime methiodide or 2-PAM. This compound is outstanding in itsability to reactivate, in vitro, oholinesterase which has been inhibitedby, for example, GB. Thus, even as compared to the very closely related4-PAM, disclosed in Example ll of the Wilson patent, 2-PAM shows muchgreater activity. With isopropyl methylphosphonylatedacetylocholinesterase the rate constant at pH 7.4 and 25 C. in thepresence of 10- M a'cetylocholine is 2x10 per mole per minute for Z-PAMand 1.4 X10 per mole per minute for 4-PA1M. The Wilson patent shows a'high rate of survival in mice which have been poisoned with paraoxon andthen treated witha) present the recommended remedy. Recently, 2-PAM hasbeen reported to enhance considerably the activity of atropine in thechemotherapeusis of poisoning due to organophosphorus compounds.

The compounds of Formula I above in which R contains from 2 to 6 carbonatoms are appreciably more effective than 2-PAM as reactivators ofGB-inhibited acetylcholinesterase and also in enhancing the activity ofatropine in both therapy and propylaxis. When If is bromide, thevariation of the rate constant for the in vitro reactivation of GBinhibited eel acetylcholinesterase at pH 7.4 and 25 C. was found to beas follows:

TABLE 1 R Rate constant (l/molcs/minutes) (cum 7 10 (cum 6x10 HM 6x10 (CM 1x10 (cum 6x10 When administered in combination with atropine toanimals poisoned with GB the order of effectiveness was somewhatdiiferent. Under these conditions the compound in which R= (CH i.e.,1,1-trin1ethy1ene bis (4- formylphyridinurn) bromide dioxime also knownas TMB-4, was most effective. In rats challenged with a 2 LD dose of GBadministered intravenously, all of a group of six animals survived ifthe atropine-TMB-4 combination was administered intravenouslyimmediately after poisoning. The atropine-2PAM combination saved onlytwo of the group of animals. On the other hand, with dogs which weregiven a 20 LD dose of GB subcutaneously the survival ratios were thesame (4/5) for the two treatments, which were given intravenously whensymptoms appeared. However, the recovery time was much shorter for thesurviving animals which received the TMB-4, i.e., 2 hours, as against 24hours for those receiving the 2-PAM.

A summary of the reactivation rates and survival ratios for thesecompounds when administered therapeutically to rats together withatropine is as follows:

TABLE 2 Reactivation rate constant Survival ratio (GB) 7X10 6X10 6X101x10 6x10 These compounds constitute our presently preferred group.

Our compounds may be employed prophylactically, i.e., injected beforeexposure to the anticholinesterase agent, e.g., GB, or therapeutically,i.e., injected subsequent to exposure.

The following series of experiments compares the efiec tiveness of ourpresently preferred compound, TMB-4,

To minimize absorption effects both the GB and TMB-4 were ordinarilygiven intravenously. However, in the therapeutic tests on rabbits, dogsand cats, the GB was administered subcutaneously, since death from 20 LDintravenous dose of GB occurs so quickly that it is virtually impossibleto give timely administration of. the antidote.

Atropine, when administered, was included in the following amounts.

Mgjkg. Rats 4 Rabbits 2 Dogs and cats 0.5

The prophylactic doses were given within two minutes prior to theinjection of the GB, the therapeutic doses so soon as poisoning symptomswere visible.

Table 3 shows the results.

TABLE 3 A. PROPHYLAOTIC The recovery periods, i.e., time fordisappearance of sypmtorns of poisoning, among survivors in the abovetests, with atropine, were as follows.

TABLE 4 2-PAM TMB-4 Animals Prophylactic Therapeutic ProphylacticTherapeutic The compounds of Formula I in which R contains from 7 to 10carbon atoms are less effective than those of our preferred group. Forthese compounds the reactivation rate constant and the survival ratiofor rats (measured as given above) were as follows, X- being bromide.

TABLE 5 R Rate Survival ratio (GB) While these compounds wereineffective in vivo against GB, they were, together with 2-PAM, veryefiective against certain other anticholinesterases, particularly thatdesignated as VX by the U.S. Army Chemical Corps. All these compoundscaused survival of all animals (survival rates of 4/ 4 and 6/6) whenadministered therapeutically to rats challenged by 2 LD doses of VX.

The compounds of Formula II exhibited properties intermediate those ofthe two subgroups of Formula I. When X- was chloride the compound hadthe following properties. (In this and all following tables the survivalratios are those for rats challenged by 2 L13 doses of GB or (VX) andthe oxime was employed therapeutically.)

Compounds of Formula III showed reactivation rates very close to thoseof our preferred group. Thus when X'- in Formula III is bromide thereactivation rate constant was 8 10 as compared to the value for the R=(CH member of our preferred group of 6x10 For the unsaturated member(Formula III) the survival ratio for rats challenged by GB was only 1/ 4as compared to 6/6 for the saturated analogue (Formula I). Both gavecomplete survival (ratios of 4/4 and 6/6) for animals challenged by VX,however.

Compounds of Formula IVa showed anomalous properties.

They gave reactivation rates which were low, but survival ratios whichwere high as compared to Z-PAM, as

shown by Table 6, X- being bromide.

TABLE 6 Survival ratio (GB) Reactivation rate constant a. 5x10 4/4 4.2x10 3 4 (C 2): HM

TABLE 7 Reactivation rate constant Survival R"EN- ratio (GB) 0,115)iN- 1. 2x10 4/4 The compounds of Formulas VI, VII and VIII, while beingof difierent structure are alike in exhibiting reactivation rateconstants which are very low as compared to Z-PAM but giving highsurvival ratios as shown by Table 8, X- being bromide in each case.

TABLE 8 Survival ratio (GB) Formula No. Reactivation rate 69 4/Negligible 4/4 67 4/4 Preparation of Compounds 4-pyridinecarboxaldehydeoxime was prepared by war-ming on a steam bath a neutralized aqueoussolution of 4-pyridinecarboxaldehyde and hydroxylamine hydrochloride.The oxime had a melting point of 130-1305 C. The 2- and 3-oximes wereproduced by similar methods.

The quaternization to produce dioximes was carried out by reacting theproper oxime with a 1, n dihaloalkane, (CH X employing a 3 :1 molarratio of oxime to halide. The unsymmetrical quaternary monoximes wereobtained by reacting the pyridine oxime with the appropriateomega-halopropyl quaternary salt in a 1.5:1 molar ratio. Two procedureswere utilized.

Procedure A: A mixture of the pyridine oxime and halide was dissolved insufficient ethanol and refluxed for the period of time specified inTable 9.

Procedure B: A mixture of the oxime and halide was dissolved in about100 ml. of ethanol and heated in a 200 ml. capped pressure bottle(carbonated beverage type) for the length of time specified. Thereaction mixtures were cooled to room temperature and the productremoved by filtration. In several instances it was necessary to addabsolute ether to eifect complete precipitation. The products wererecrystallized from ether. This procedure was usually employed becauseof its simplicity.

Table 9 gives the procedure, yields and melting points forrepresentative compounds.

TABLE 9 For- Substituents Melting (n1) mula Gondi- Yield or decom- No.tions percent position (d) Halide R R point, C.

I Br 35. 0 300 m. I Br 88. 2 238-241 (I. I Br 81.0 239-241 (I. I Br 95.0 208-210 (1. I Br 85. 0 219-223 6.

II... 01 B, as hr 70 300 m. IVa Br (011m B, 60 r 68 208-211 m. IVai. Br(Slim B, 60 hr 226-231 m. 1%... Br B, 20 111:. 83.5 248-251 in. V Br(021393 B, 69 11L- 43 230-231 (1. V Br pyridine B, 64 11L- 10 223-226(1.

mg. VIII." Br B, hr 16 201-203 (1.

Further details regarding the preparation and properties of certain ofour compounds are given in the following publications by us and ourassociates:

Pyridine Alcloximes, by Edward J. Poziomek, Bernnie E. Hackley, Jr., andGeorge M. Steinberg, Journal of Organic Chemistry, vol. 23, pp. 714-717(May 1958); and Chemotherapeutic Effectiveness of T rimethylene Bis(4-Formy1 Pyridinium Bromide) Dioxime in Anticholinesterase Poisoning,by Edmund Bay, S. Kropp and L. F. Yates, Proceedings of the Society forExperimental Biology and Medicine, vol. 98, pages 107-109 (May 1958).These articles are to be considered incorporated by reference in thisspecification.

While we have shown a number of specific examples of compounds and theiruse, it will be obvious that various changes can be made withoutdeparting from our invention, which is defined by the following claim.

We claim:

A method of therapeutically treating a mammal which has been poisoned bya compound having high anticholinesterase activity which comprisesinjecting a composition comprising atropine and 1,1'-(2-butenylene)bis(4- formylpyridinium) halide dioxime.

' References Cited in the file of this patent Poziomek: Am. Chem. Soc.Abst. of Papers, 132nd Meeting, 1957, pages 16-0.

